Blood cells and some hematological parameters of red drum (Linnaeus, 1766) in Vietnam / Células sanguíneas e alguns parâmetros hematológicos do tambor vermelho (Linnaeus, 1766) no Vietnã

This research focuses on hematological characteristics, erythrocyte morphology and some biochemical parameters of red drum Sciaenops ocellatus (Perciformes: Sciaenidae), cultured in natural water environment in areas near river mouth (L1), estuaries (L2) in Ha Tinh province and coastal areas (L3) in Nha Trang city, Khanh Hoa province of Vietnam. A total of 18 speciments were examined in research, six in each location. Blood was drawn from the tail vein, using a microscope to research morphology and automated gauges to determine blood biochemical parameters. Analysis of blood samples showed that the rate of red drum's erythrocyte morphology disorders in all three locations was quite high. The two main types of disorders were nuclear deformity and nuclear-matter distribution. Changes in erythrocyte size, shape and nuclear were related to salt concentration at culture locations. Blood hemoglobin content was stable in all three regions. Other hematological parameters such as the number of erythrocytes, blood biochemical parameters (glucose, SGOT, SGPT, urea, creatine, plasma iron, albumin, and protein) have differences among the locations, which showed the different reactions of the same species with different environmental conditions.

Esta pesquisa se concentra nas características hematológicas, na morfologia eritrocitária e em alguns parâmetros bioquímicos do tambor vermelho Sciaenops ocellatus (Perciformes: Sciaenidae), cultivado em ambiente aquático natural em áreas próximas à foz do rio (L1), em estuários (L2) na província de Ha Tinh e em áreas costeiras (L3) na cidade de Nha Trang, província de Khanh Hoa do Vietnã. Um total de 18 espécimes foi examinado na pesquisa, seis em cada local. O sangue foi coletado da veia da cauda, usando um microscópio para pesquisar a morfologia e medidores automatizados para determinar os parâmetros bioquímicos do sangue. A análise das amostras de sangue mostrou que a taxa de distúrbios morfológicos dos eritrócitos do tambor vermelho em todos os três locais era bastante alta. Os dois principais tipos de distúrbios eram deformidade nuclear e distribuição de matéria nuclear. Mudanças no tamanho, na forma e no núcleo dos eritrócitos foram relacionadas à concentração de sal nos locais de cultivo. O conteúdo de hemoglobina no sangue era estável em todas as três regiões. Outros parâmetros hematológicos, como número de eritrócitos, parâmetros bioquímicos sanguíneos (glicose, SGOT, SGPT, ureia, creatina, ferro plasmático, albumina e proteína), apresentam diferenças entre os locais, o que evidencia diferentes reações de uma mesma espécie em diferentes ambientes e condições.

Avanços e perspectivas no diagnóstico molecular da leucemia mieloide aguda: revisão sistemática / Advances and perspectives in the molecular diagnosis of acute myeloid leukemia: systematic review

O câncer do colo do útero é o quarto tumor mais comum entre mulheres no mundo e o terceiro no Brasil. A detecção precoce e a identificação das lesões cervicais são essenciais no rastreamento do câncer cervical. Nos últimos anos, vários marcadores têm sido apresentados como possíveis candidatos para a triagem eficiente de exames citológicos com anormalidades das células epiteliais. O objetivo deste trabalho é analisar a correlação com a expressão imuno-histoquimica dos biomarcadores p16 e Ki-67 com lesão intraepitelial cervical de alto grau na detecção molecular DNA/HPV de alto risco. A metodologia de pesquisa empregada é uma revisão sistemática, realizada por meio de buscas nas bases de dados eletrônica Literatura Latino-Americana em Ciência e Saúde (LILACS), Scientific Electronic Library Online (SCIELO), National Library of Medicine (MEDLINE) de artigos publicados no período de 2005 a 2019 nos idiomas português, inglês e espanhol. Concluiu-se que o uso das proteínas p16 e Ki67 auxilia na identificação das mudanças que acontecem durante a progressão da lesão cervical, aprimorando os métodos de rastreio atuais. O gene p53, a pRb e ciclinas também têm um papel crítico na carcinogênese e, desta maneira, também têm sido indicados para entrar nos painéis de estudo.

Introduction: Acute myeloid leukemia (AML) is a genetically heterogeneous malignant clonal disorder of the hematopoietic system, characterized by the uncontrolled proliferation of abnormal and immature blast cells and impaired production of normal blood cells. Objective: to present the advances in diagnosis in Acute Myeloid Leukemia through the techniques of molecular biology. Methodology: The research was carried out in the electronic databases for scientific articles of the search portal PubMed, NCBI, BVSMS, also SciELO. Results: Molecular techniques, including real-time quantitative PCR (RT-qPCR), digital PCR and technologies based on new generation sequencing, can be standardized and used to detect AML-associated gene changes. Conclusion: The advance in molecular diagnosis can be promising in the ideal and personalized treatment.

Decreased levels of cathepsin Z mRNA expressed by immune blood cells: diagnostic and prognostic implications in prostate cancer

Cathepsin Z (CTSZ) is a cysteine protease responsible for the adhesion and migration of both immune and tumor cells. Due to its dual role, we hypothesized that the site of CTSZ expression could be determinant of the pro- or anti-tumorigenic effects of this enzyme. To test this hypothesis, we analyzed CTSZ expression data in healthy and tumor tissues by bioinformatics and evaluated the expression levels of CTSZ mRNA in the blood cells of prostate cancer (PCa) patients by qRT-PCR compared with healthy subjects, evaluating its diagnostic and prognostic implications for this type of cancer. Immune cells present in the blood of healthy patients overexpress CTSZ. In PCa, we found decreased CTSZ mRNA levels in blood cells, 75% lower than in healthy subjects, that diminished even more during biochemical relapse. CTSZ mRNA in the blood cells had an area under the curve for PCa diagnosis of 0.832, with a 93.3% specificity, and a positive likelihood ratio of 9.4. The site of CTSZ mRNA expression is fundamental to determine its final role as a protective determinant in PCa, such as CTSZ mRNA in the blood cells, or a malignant determinant, such as found for CTSZ expressed in high levels by different types of primary and metastatic tumors. Low CTSZ mRNA expression in the total blood is a possible PCa marker complementary to prostate-specific antigen (PSA) for biopsy decisions, with the potential to eliminate unnecessary biopsies.

Comparative study on erythrogram parameters of Cell-Dyn Ruby and Humacount 5l / Estudio comparativo de los parámetros del eritrograma de Cell-Dyn Ruby y Humacount 5l

SUMMARY: The use of hematological counts for the prevention, diagnosis and follow-up of hematological diseases has increased. Indeed, the correct operation of a clinical laboratory is essential to producing comparable results. However, there is a paucity of validation and reproducibility studies among the different existing methods for clinical analysis. Therefore, our aim was to assess the commutability of the results provided by analyzers with different measuring systems. Sixty venous blood samples were obtained from patients, without discriminating for age or sex. Then, an automated hematological analysis was performed using the Cell-Dyn Ruby and HumaCount 5L instruments. The variables measured were: RBC, Hb, HCT, MCV, MCH and MCHC. The data were compared by a one-way ANOVA and Pearson's correlation coefficient. Statistical significance was fixed at p < 0.05. There were no statistically significant differences for RBC, HCT, MCH or MCHC. In addition, with the exception of MCHC, all the analytes showed a good correlation coefficient between the two instruments. There is a variety of automated systems for the clinical laboratory and it is essential for the clinician to know the different methodologies used in hematological analyzers as well as their sensitivity and specificity. Therefore, our results are useful for demonstrating the importance of practical knowledge of the analyzers mentioned.

RESUMEN: El uso de recuentos de células sanguíneas para la prevención, diagnóstico y monitoreo de enfermedades hematológicas ha ido en aumento. Por ello, el funcionamiento correcto de un laboratorio clínico es indispensable para producir resultados comparables. Sin embargo, existen pocos estudios de validación y reproducibilidad de los diferentes métodos de análisis clínico existentes. Por lo tanto, nuestro objetivo fue evaluar la intercambiabilidad de los resultados entregados por los analizadores que utilizan diferentes sistemas de medición. Se obtuvieron sesenta muestras de sangre venosa de pacientes, sin discriminar por edad o sexo. Los eritrogramas fueron obtenidos utilizando los analizadores automatizados Cell-Dyn Ruby y HumaCount 5L. Las variables medidas fueron: RBC, Hb, HCT, MCV, MCH y MCHC. Los datos fueron comparados por ANOVA a una vía y la correlación de Pearson. La significación estadística se estableció en el nivel estándar p<0,05. No hubo diferencias estadísticamente sig- nificativas para RBC, HCT, MCH y MCHC. Con la excepción de la MCHC, todos los analitos presentaron un buen coeficiente de correlación entre los dos analizadores comparados. Existen varios sistemas de automatización para su uso en laboratorios clínicos. Por lo tanto, es primordial para el clínico estar familiarizado con las diferentes metodologías utilizadas en los analizadores de sangre, así como su sensibilidad y especificidad. Nuestros resultados son útiles para mostrar la importancia del conocimiento práctico de los diferentes sistemas de medidas comparados.

Comparación de parámetros hematológicos y morfología celular en muestras de sangre con EDTA K2 y EDTA K3 / Comparison of hematological parameters and cellular morphology in blood samples with EDTA K2 and EDTA K3

El EDTA es el anticoagulante de elección en los laboratorios de hematología para la conservación de la muestra de sangre total. Existen dos tipos, EDTA K2 y EDTA K3, y su diferencia radica en la cantidad de moléculas de potasio. Algunas guías sugieren que hay diferencias entre el anticoagulante EDTA K2 y el K3 para el proceso del hemograma; sin embargo, con las nuevas presentaciones de los tubos que traen las casas comerciales, no se tiene claro si en realidad aún hay diferencia entre los dos anticoagulantes, y si esto puede alterar el resultado del hemograma, tanto en el resultado cuantitativo, como en el cualitativo. Objetivo. Comparar los recuentos leucocitarios, la hemoglobina, el hematocrito, el volumen corpuscular medio, las plaquetas y la morfología celular en muestras de sangre periférica con EDTA K2 y EDTA K3, en diferentes tiempos (0, 1 y 2 horas). Materiales y métodos. Se realizó un estudio cuasi-experimental, multivariado, multifactorial, que tiene como unidad de análisis la sangre anticoagulada con EDTA K2 y EDTA K3, extraída de 53 individuos a través de un muestreo no probabilístico por conveniencia. Resultados. Al comparar los resultados del estudio morfológico por medio del extendido de sangre periférica y los datos cuantitativos del hemograma, se encontró que no hay diferencias estadísticamente significativas usando EDTA K2 o K3. Conclusión. Se evidenció que el uso del EDTA K2 o EDTA K3 como anticoagulante de elección, procesando las muestras en un tiempo adecuado después de su recolección, no afecta los parámetros cuantitativos del hemograma automatizado ni los morfológicos.

EDTA is the anticoagulant of choice in hematology laboratories for the conservation of whole blood samples. There are two types, K2 EDTA and K3 EDTA, and their difference lies in the amount of potassium molecules. Some guidelines suggest that there are differences between K2 and K3 EDTA for the blood analysis process. However, with the new collection tubes offered by the commercial suppliers, it is not clear if in fact there is a difference between the two anticoagulants that would result in changes in blood parameters and cell morphology. Objective. To compare leukocyte counts, hemoglobin, hematocrit, mean corpuscular volume, platelets and cell morphology in peripheral blood samples collected with K2 EDTA and K3 EDTA, at different times (0, 1 and 2 hours). Materials and methods. A quasi-experimental, multivariate, multifactorial study was carried out, with anticoagulated blood as the unit of analysis, either with K2 EDTA or K3 EDTA, extracted from 53 subjects through a non-probabilistic sampling for convenience. Results. There was no statistically significant difference when comparing results of the peripheral blood smear and the quantitative hematological parameters using K2 or K3 EDTA. Conclusion. The use of either K2 EDTA or K3 EDTA as the anticoagulant of choice, when processing samples within a suitable time after their collection, proved equally satisfactory for both quantitative and morphological parameters

Enfermedad de Gaucher: caso clínico / Gaucher's disease: a clinical case

La enfermedad de Gaucher es una patología de depósito lisosomal, autosómica recesiva, con mutación en el gen GBA, que afecta principalmente al hígado, bazo, huesos y a la médula ósea. Es una enfermedad rara con baja incidencia mundial. Existen 3 tipos; el tipo I es el más frecuente en la población pediátrica, y el tipo III es el de peor pronóstico y presenta manifestaciones neurológicas. El diagnóstico se realiza con el tamizaje prenatal de enfermedades de depósito, más específicamente de mutaciones GBA. El diagnóstico definitivo se realiza al detectar ß-glucocerebrosidasa, y los estudios genéticos para la tipificación del gen afectado: en el tipo 1 mutación N370S y mutaciones L444P o D409H en tipo 2 y 3. El tratamiento se realiza con terapia de reemplazo enzimático o con la terapia de reducción de sustrato. Describimos en este artículo a un paciente preescolar del Hospital Enrique Garcés, con cuadro respiratorio, hepato-esplenomegalia y afectación de las tres líneas celulares sanguíneas.

Gaucher's disease is an autosomal recessive lysosomal storage pathology, with mutation in the GBA gene, which mostly affects the liver, spleen, bone and bone marrow. It is a rare disease with low global incidence. There are 3 types: type I is the most frequent in the pediatric population, and type III has the worst prognosis and presents neurological manifestations. Diagnosis is made with prenatal screening of storage diseases, and more specifically GBA1 mutations. The definitive diagnosis is made by detecting ß-glucocerebrosidase, and genetic studies for the detarmination of the affected gene, in type 1 mutation N370S and L444P mutations or D409H in type 2 and 3. The treatment is enzyme replacement therapy or substrate reduction therapy. This article describes a preschool patient of Enrique Garcés Hospital, with respiratory symptoms, hepato-splenomegaly and involvement of the three blood cell lines.

Blood cells profile in umbilical cord of late preterm and term newborns / Perfil de hemograma em sangue de cordão umbilical de recém-nascidos pré-termo tardio e a termo

ABSTRACT Objective: To describe the hematological profile in cord blood of late preterm and term newborns and compare blood indices according to sex, weight for gestational age and type of delivery. Methods: Cross-sectional study with late preterm and term newborns in a second-level maternity. Multiple gestation, chorioamnionitis, maternal or fetal hemorrhage, suspected congenital infection, 5-minute Apgar <6, congenital malformations, and Rh hemolytic disease were excluded. Percentiles 3, 5,10, 25, 50, 75, 90, 95 and 97 of blood indices were calculated for both groups. Results: 2,662 newborns were included in the sample, 51.1% males, 7.3% late preterms, 7.8% small for gestational age (SGA) and 81.2% adequate for gestational age (AGA). Mean gestational age was 35.6±1.9 and 39.3±1.0 weeks, respectively, for premature and term neonates. The erythrocytes indices and white blood cells increased from 34-36.9 to 37-41.9 weeks. Basophils and platelets remained constant during gestation. Premature neonates presented lower values ​​of all blood cells, except for lymphocytes and eosinophils. SGA neonates presented higher values ​​of hemoglobin, hematocrit and lower values of leukocytes, neutrophils, bands, segmented, eosinophils, monocytes and platelets. Male neonates presented similar values ​​of erythrocytes and hemoglobin and lower leukocytes, neutrophils, segmented and platelets. Neonates delivered by C-section had lower values ​​of red blood cells and platelets. Chronic or gestational hypertension induced lower number of platelets. Conclusions: Blood cells increased during gestation, except for platelets and basophils. SGA neonates had higher hemoglobin and hematocrit values and lower leukocytes. Number of platelets was smaller in male SGAs, born by C-section and whose mothers had hypertension.

RESUMO Objetivo: Descrever o perfil hematológico em sangue de cordão de recém-nascidos pré-termo tardio e a termo e comparar parâmetros hematimétricos segundo sexo, adequação peso idade gestacional e tipo de parto. Métodos: Estudo transversal com recém-nascidos pré-termo tardio e a termo, em maternidade de nível secundário. Excluíram-se gestação múltipla, corioamnionite, hemorragia materna ou fetal, suspeita de infecção congênita, Apgar no 5o minuto <6, malformações congênitas e doença hemolítica Rh. Calcularam-se os percentis 3, 5, 10, 25, 50, 75, 90, 95 e 97 dos parâmetros hematológicos. Resultados: Incluíram-se 2.662 recém-nascidos, 51,1% do sexo masculino, 7,3% prematuros tardios, 7,8% pequenos para a idade gestacional e 81,2% adequados. A idade gestacional foi 35,6±1,9 e 39,3±1,0 semanas, respectivamente, nos prematuros e termos. As séries vermelha e branca aumentaram de 34-36,9 para 37-41,9 semanas, exceto basófilos e plaquetas, que permaneceram constantes. Os prematuros apresentaram menores médias nas séries vermelha, plaquetária e branca, com exceção de linfócitos e eosinófilos. Recém-nascidos pequenos para a idade gestacional apresentaram maiores valores de hemoglobina e hematócrito e menores de leucócitos, neutrófilos, bastonetes segmentados, eosinófilos, monócitos e plaquetas. Recém-nascidos masculinos apresentaram taxas semelhantes de hemoglobina e hematócrito e menores de leucócitos, neutrófilos, segmentados e plaquetas. Na cesárea, as células vermelhas e as plaquetas foram menores que no parto vaginal. O número de plaquetas foi menor na hipertensão crônica ou gestacional. Conclusões: As células sanguíneas aumentaram durante a gestação, exceto plaquetas e basófilos. Recém-nascidos pequenos para a idade gestacional apresentaram maiores taxas de hemoglobina e hematócrito e menores de células brancas. O número de plaquetas foi menor no recém-nascido pequeno para a idade gestacional, masculino, nascido por cesárea e de mãe hipertensa.

Use of Inert Gases for the Preservation of Nuclear Blood Cells

Abstract The subject of the study was the stability of human white blood cell membranes subject to noble gases (xenon ad krypton, 0.6 mPa) clathrate cryoanabiosis (‒80°C). A unique portable stainless steel low pressure container with a compartment for flexible plastic container was designed to ensure that the cells are saturated with gases. The samples were warmed after 1 and 30 days in a water bath (+38°C) for 35-50 sec, while the container was being tilted (2-3 times per second), until the temperature of the biological object reached +3±1°C. It was demonstrated that after 30 days of clathrate anabiosis (-80°C) over 95% (of the original number) of leukocytes remain viable, and cell membranes of 54.5±3.4% of them is resistant to trypan blue; granulocyte survival rate is 73.5±2.7%, original lipid peroxidation rate and antioxidant activity are retained. Biological object cryopreservation in noble gases environment is a promising trend in biology and medicine.

Avaliação dos efeitos da fibrina rica em plaquetas (FRP) comparada a um anti-inflamatório não esteroidal (AINE) na resposta inflamatória e reparadora de defeitos críticos em calota de ratos / Evaluation of the effects of Platelet Rich Fibrin (PRF) compared to an Non-Steroidal (NSAID) in the inflammatory and repair of critical defects in rat's calvaria

O objetivo deste estudo foi avaliar a influência da Fibrina Rica em Plaqueta (FRP) no processo inflamatório em defeitos críticos em calotas de ratos e sua consequente reparação tecidual. Foram utilizados 128 (cento e vinte e oito) ratos (Rattus norvegicus, albinus, Wistar), adultos, com peso corporal entre 450 e 500g. Os animais foram divididos aleatoriamente em 4 grupos equitativos que compuseram a amostra do trabalho, onde no grupo coágulo (GC) foi realizado o defeito ósseo de tamanho crítico preenchido com coágulo sanguíneo; grupo anti-inflamatório não esteroidal (AINE) que teve os defeitos de tamanho crítico preenchidos com coágulo sanguíneo e administrado cetoprofeno (10mg/kg dia); o grupo fibrina rica em plaquetas (FRP) com defeitos de tamanho crítico preenchidos com preparado de fibrina rica em plaquetas autóloga; e o grupo fibrina rica em plaquetas mais AINE (FRP + AINE) com defeitos de tamanho crítico preenchidos com preparado de fibrina rica em plaquetas autóloga e administrado cetoprofeno (10mg/kg dia). Cada grupo foi avaliado nos períodos de 2, 7, 14 e 28 dias e os espécimes analisados através da histometria, micro-CT e teste ELISA para presença de TNFα. Os dados quantitativos foram submetidos aos testes estatísticos ANOVA 1/2 fatores ou Kruskal-Wallis e pos Tukey e Dunn (p<0,01). Os resultados histométricos e microtomográficos evidenciaram maior formação óssea para o grupo PRF em comparação aos demais grupos (p<0,05) e menor presença de TNF-alfa no período inicial no grupo PRF comparado ao grupo controle (p<0,05). Conclui-se que o PRF foi favorável desde os períodos iniciais até os mas tardios, auxiliando na resposta inflamatória e neoformação óssea(AU)

The objective of this study was to evaluate the influence of Platelet Rich Fibrin (PRF) on the inflammatory process in critical defects in rat calvaria and its consequent tissue repair. One hundred twenty-eight adult rats (Rattus norvegicus, albinus, Wistar) with body weight between 450 and 500g were used for the study. The animals were randomly divided into 4 equitable groups that composed the work sample. In the clot group (GC) the critical size defect was filled with blood clot; the non-steroidal anti-inflammatory group (NSAID) had the critical size defects filled with blood clot and ketoprofen was given pos-operatively (10mg / kg day); the platelet-rich fibrin group (FRP) had the critical-size defects filled with autologous platelet-rich fibrin preparation; and platelet-rich fibrina group plus NSAIDs (FRP + NSAID) had the critical-size defects filled with autologous platelet-rich fibrin preparation and administered ketoprofen post-operatively (10mg / kg day). Each group was evaluated at 2, 7, 14 and 28 days after the surgical procedures. The samples were evaluated through histometry, micro-CT and ELISA for the presence of TNFα. The quantitative data were submitted to the statistical test ANOVA 1/2 factors or Kruskal-Wallis and post Tukey and Dunn (p <0.01). The histometric and microtomographic results showed higher bone formation for the PRF group compared to the other groups (p <0.05) and lower TNF-alpha in the initial period in the PRF group compared to the control group (p <0.05). It was concluded that the PRF was favorable since the beginning through the later periods, aiding in the inflammatory response and bone neoformation(AU)

Extractos vegetales de tres especies del género Baccharis inducen la proliferación de células mononucleares humanas / Plant extracts of three species of the genus Baccharis induce human mononuclear cells proliferation

Las plantas de uso en medicina tradicional constituyen una fuente importante de compuestos con actividad inmunomoduladora; entre ellas las especies del género Baccharis, conocidas popularmente como "Jaguareteka´a" en nuestro país, son ampliamente empleadas. En este estudio se evaluó la actividad inmunomoduladora de extractos metanólicos de tres especies del género Baccharis (B. trimera, B. notosergilay B. punctulata) sobre la proliferación de células mononucleares humanas de sangre periférica. Los extractos de las tres especies estudiadas estimularon la proliferación de las células mononucleares. Específicamente, el extracto de B. notosergila estimuló la proliferación celular a todas las concentraciones probadas (5, 10, 25 y 50 µg/mL), mientras que los extractos de B. trimera y B. punctulata mostraron este efecto a 5 y 10 µg/mL. Además, por presentar mayor inducción de la proliferación, se realizó un fraccionamiento con diferentes solventes del extracto metanólico de B. notosergila y B. punctulata. La fracción de acetato de etilo de ambos extractos vegetales aumentó la proliferación celular, sugiriendo que compuestos de polaridad media son los responsables de esta actividad. Estos resultados demuestran que los extractos de B. trimera, B. notosergila y B. punctulata poseen actividad inmunomoduladora sobre células mononucleares humanas y servirán de base a otros estudios para determinar el o los componentes activos de los extractos sobre el sistema inmune(AU)

Plants used in traditional medicine are an important source of compounds with immunomodulatory activity. Species of the genus Baccharis, popularly known as "Jaguareteka'a" in our country, are used in folk medicine for the treatment of liver, gastrointestinal, inflammatory and infectious diseases. In this study, we evaluated the immunomodulatory activity of methanolic extracts of three species of the genus Baccharis (B. trimera, B. notosergila and B. punctulata) on the proliferation of human peripheral blood mononuclear cells. Extracts of the three species studied stimulated the proliferation of mononuclear cells. The extract of B. notosergila stimulated cell proliferation at all concentrations tested, while extracts of B. trimera and B. punctulata stimulated at 5 and 10 µg/mL. In addition, we carried out a separation with different solvents of the methanolic extract of B. notosergila and B. punctulata. The ethyl acetate fraction of both plant extracts induced the proliferation of immune cells. These results show that the extracts of B. trimera, B. notosergila and B. punctulata had immunomodulatory activity on human mononuclear cells. Future work will be required to identify the components responsible for the activity on the immune system(AU)

Assessment of hematological parameters in workers exposed to organophosphorus pesticides, carbamates and pyrethroids in Cundinamarca 2016-2017 / Evaluación de parámetros hematológicos en trabajadores expuestos a pesticidas organofosforados, carbamatos y piretroides, Cundinamarca 2016-2017

ABSTRACT Objective To establish hematotoxic alterations through clinical and paraclinical exploration in workers who are exposed to organophosphorus pesticides, carbamates and pyrethroids (OPCP) due to their work in production, packaging, distribution and fumigation processes in Cundinamarca-Colombia between 2016 and 2017. Materials and Methods A cross-sectional descriptive epidemiological study was carried out on a sample of 92 workers from six companies, mostly aged between 18 and 30 years, of which 61 % were males and 39 % females, and 71 % were workers in the operational area and 29 % in the administrative area. Univariate, bivariate and multivariate analyses were performed. Results Clinical exploration reported findings in 17 % of the sample group, of which only 2 % presented with erythrocyte cholinesterase outside the reference range. The values of hematological parameters such as peripheral blood smear (PBS) and complete blood count (CBC) were outside the range in 15 % and 47 % of the sample, respectively. Discussion The results suggest that there are hematological alterations in this group that could possibly be associated with chronic exposure to OPCP.

RESUMEN Objetivo Determinar las alteraciones hematotóxicas a través de una exploración clínica y paraclínica, en trabajadores que por su oficio se exponen a pesticidas organofosforados, carbamatos y piretroides (POCP), en procesos de producción, envase, distribución y fumigación, en el departamento de Cundinamarca Colombia. Metodología Se realizó un estudio epidemiológico descriptivo de corte transversal. Se realizó un análisis univariado, bivariado y multivariado. La muestra se conforma por 92 trabajadores de seis empresas, la mayoría entre los 18 y 30 años, de los cuales el 61 % son hombres y el 39 % mujeres, 71 % se desempeñan en el área operativa y 29 % en el área administrativa. Resultados Es de resaltar que se tienen hallazgos en la exploración clínica en el 17 % del grupo participante, solo el 2 % presenta la colinesterasa eritrocitaria por fuera del rango de referencia. Los parámetros hematológicos como el frotis de sangre periférica (FSP) presentan valores por fuera de los rangos en el 15 % y el cuadro hemático (CH) tiene valores por fuera de los rangos en el 47 % de las personas. Discusión Los resultados sugieren que existen alteraciones hematológicas en este grupo y que posiblemente podrían estar asociadas con la exposición crónica a POCP.

Aspectos morfológicos e ultraestruturais de células sanguíneas de Crotalus durissus terrificus / Morphological and ultrastructural aspects of Crotalus durissus terrificus blood cells

A avaliação hematológica, de importância comprovada como um meio auxiliar de diagnóstico ao clínico de pequenos animais domésticos, vem se tornando comum em animais selvagens não apenas para a clínica, mas para a avaliação do manejo e como estudo auxiliar para a fisiologia das várias espécies. Tendo em vista o aumento da demanda para a produção de várias drogas de importância farmacêutica, a criação de serpentes peçonhentas vem se tornando comum a ponto destes animais já serem reconhecidos como sendo de produção. O conhecimento do manejo e da clínica destes animais ainda é escasso e a mortalidade é elevada nos criatórios, tornando urgente a ampliação destes. Embora alguns estudos hematológicos já tenham sido realizados em cascavéis (Crotalus durissus) os dados analisados ainda são insipientes, notadamente em relação à caracterização das células do sangue e poucos estudos em microscopia eletrônica foram realizados em serpentes. Com o objetivo de caracterizar as células sanguíneas morfologicamente, sob microscopia óptica e ultraestrutural, foram coletadas amostras de sangue de 52 de indivíduos da subespécie Crotalus durissus terrificus para a realização de esfregaços sanguíneos e avaliação ultraestrutural. Concluiu-se que a coloração hematológica de Giemsa permite a avaliação morfológica e a diferenciação das células sanguíneas em serpentes assim como a visualização de hemoparasitos. A avaliação ultraestrutural permite evidenciar as organelas celulares e a diferenciação entre as células, inclusive entre os tipos leucocitários, porém ainda são necessários outros estudos para que seja elucidada a hipótese da existência dos eosinófilos na espécie estudada assim como é necessária melhor caracterização dos grânulos dos azurófilos para que se confirme uma possível diferença entre os monócitos típicos e os azurófilos.(AU)

Hematological evaluation, important for the diagnostic by the small domestic animal clinician, has become common in wildlife clinic, and for handling and study of the physiology of various species. Given the increased demand for drug production of pharmaceutical importance, the breeding of venomous snakes has become common and is already recognized as production. Knowledge of the management and clinics of snakes is still insufficient and their mortality is high. Although some hematological studies have already been conducted in the rattlesnake (Crotalus durissus), the analyzed data are still insufficient, especially with respect to the characterization of blood cells, and few electron microscopy studies have been performed on snakes. In order to characterize morphologically blood cells with light and ultrastructural microscopy, blood samples from 52 individuals of subspecies of Crotalus durissus terrificus were collected to perform blood smears and ultrastructural evaluation. It was concluded that hematologic Giemsa staining allows morphological evaluation and differentiation of the blood cells as well as of snake hemoparasites. The ultrastructural evaluation will highlight the cell organelles and differentiation between cells, including leukocyte types; although still further studies are needed to elucidate the hypothesis of eosinophils in the species studied as also is necessary a better characterization of azurophilic beads to confirm a possible difference between the typical monocyte and the azurophilic.(AU)

Conjuntivitis calcárea crónica / Calcareous chronic conjunctivitis

Al hacer sistemáticamente el examen de la conjuntiva en mis enfermos, y muy especialmente en aquellos que adolecen de vicios de refracción, he observado una afección conjuntival caracterizada por la presencia de corpúsculos calcáreos incrustados en el tejido de la conjuntiva palpebral. Sintomatología. Cuando se baja el párpado inferior o se voltea el párpado superior, se ven en diversos puntos de la conjuntiva palpebral, tanto en el borde palpebral como sobre el tarso y muy rara vez en el fondo de saco, unos corpúsculos más o menos del tamaño de una cabeza de alfiler, de color blanco amarillento y de consistencia dura, petrosa, que levantan la mucosa, siendo verdaderos cuerpos extraños incrustados dentro de la misma conjuntiva.

Eriptosis: mecanismos moleculares y su implicación en la enfermedad aterotrombótica / Eryptosis: molecular mechanisms and their implications in atherothrombotic disease

Introducción: La eriptosis se ha descrito como el proceso de muerte celular programada en el eritrocito antes de la senectud. Puede desencadenarse en situaciones como: el estrés osmótico, el estrés oxidativo, la exposición a metales pesados, entre otros factores. Diversos estudios sugieren que los eritrocitos pueden desempeñar un papel activo en la hemostasia normal o anormal en ciertas condiciones en las que se produce perturbación de la membrana de estas células. Objetivo: Describir los mecanismos involucrados en la eriptosis y su estrecha relación con los procesos de adhesión a la pared vascular que conllevan a la enfermedad trombótica. Método: Se hizo una revisión narrativa a partir de la literatura encontrada en las bases de datos PubMed y Science Direct utilizando las palabras claves. Se seleccionaron 51 artículos originales, 20 revisiones de la literatura y un estudio de casos, que se ajustaban a las exigencias del objetivo. Se revisaron los resúmenes de forma separada e independiente. Seguidamente se buscaron las publicaciones en el texto completo para la revisión. Conclusiones: La eriptosis se caracteriza por la disminución del volumen celular, la vesiculación y la translocación del fosfatidil serina hacia la superficie externa de la membrana plasmática. Las alteraciones en la distribución de los fosfolípidos favorecen los procesos de adhesión celular a la pared vascular, conllevando al deterioro de la microcirculación, lo cual puede ocasionar importantes trastornos a nivel cardiovascular. La comprensión y el esclarecimiento de la eriptosis pueden ser esenciales para la búsqueda de nuevas dianas terapéuticas, encaminadas a ofrecer otras alternativas farmacológicas en el tratamiento de la enfermedad cardiovascular.

Introduction: Eryptosis has been described as programmed process of cellular death in erythrocytes before old age. It can be triggered, among other factors, by situations such as osmotic stress, oxidative stress or exposure to heavy metals. Several studies suggest that erythrocytes can play an active role in normal or abnormal haemostasis in certain conditions where the membrane of these cells is perturbed. Objetive: To describe the mechanisms involved in eryptosis and their close relationship with the processes of adhesion to the vascular wall that entail the thrombotic disease. Methods: A narrative review was carried out from the literature found in data base PubMed and Science Direct by using the key words. 51 original articles, 20 literature reviews and one case study complying with the requirements were selected. Abstracts were reviewed separately and independently. Complete publications were then located for review. Conclusions: Eryptosis is characterised by the decrease in cell volume, the vesiculation and the translocation of phosphatidylserine towards the outer surface of the plasma membrane. Disorders in the distribution of the phospholipids favour processes of cell adhesion to the vascular wall, causing impairment of microcirculation, which can result in important cardiovascular diseases. Understanding and clarifying eryptosis could be essential for finding new therapeutic targets, aimed at offering other pharmacological alternatives for the treatment of cardiovascular diseases.

Avaliação imuno-histoquímica de células de Langerhans e densidade microvascular em ameloblastomas / Immunohistochemistry assessment of Langerhans cells and microvessel density in ameloblastomas

Este estudo objetivou avaliar e correlacionar a microdensidade vascular (MDV) e a quantidade de células de Langerhans (CLs) presentes em ameloblastoma (AB). Vinte e cinco lesões em blocos parafinados de AB foram analisados através de técnica imuno-histoquímica onde foram empregados dois marcadores, anti-CD1a e anti-CD207, para quantificar as células de Langerhans, e o CD34 para avaliar a MDV. A imunomarcação para o CD1a, CD207 e CD34 foi observada em 88% dos casos analisados, mostrando uma significativa associação estatística (p = 0.001, FisherÆs test). Não foi observada correlação estatística entre MDV e CLs, nem relação entre as imunomarcações com os tipos unicístico e sólido. No entanto, a imunomarcação pelo CD1a e CD207 teve uma correlação estatisticamente significante (p valor = 0,001, teste de Spearmann). As CLs e a MDV parecem influenciar na imunopatogênese do ameloblastoma, apesar de não ter sido encontrada correlação estatisticamente significante entre esses dois achados, mas possivelmente por essas células dendríticas produzirem citocinas inflamatórias indutoras de destruição óssea e mitose celular.

The aim of this study was to assess and correlate microvessel density (MVD) and the quantity of Langerhans cells (LC) present in ameloblastomas (AB). Twenty-five paraffin-embedded blocks of AB lesions were analyzed using the immunohistochemical technique in which anti-CD1a and anti-CD207 markers were used to quantify the Langerhans cells and the CD34 marker to assess MVD. In 88% of the analyzed cases immunostaining was observed for CD1a, CD207, and CD34, with statistically significant association (p = 0.001, FisherÆs test). No statistical correlation was observed between MVD and LCs nor between immunostainings with solid and unicyst ameloblastoma. However, statistically significant correlation (p value = 0,001, Spearman test) was observed between CD1a and CD207 immunostaining. The LCs and MVD seemed to influence immunopathogenesis of ameloblastoma, although no statistically significant correlation was observed between these two findings, probably because these dendritic cells produce inflammatory cytokines that induce bone destruction and cell mitosis.

Evaluation of the effects of ischemic preconditioning on the hematological parameters of rats subjected to intestinal ischemia and reperfusion

OBJECTIVES: Intestinal ischemia/reperfusion often leads to acute lung injury and multiple organ failure. Ischemic preconditioning is protective in nature and reduces tissue injuries in animal and human models. Although hematimetric parameters are widely used as diagnostic tools, there is no report of the influence of intestinal ischemia/reperfusion and ischemic preconditioning on such parameters. We evaluated the hematological changes during ischemia/reperfusion and preconditioning in rats. METHODS: Forty healthy rats were divided into four groups: control, laparotomy, intestinal ischemia/reperfusion and ischemic preconditioning. The intestinal ischemia/reperfusion group received 45 min of superior mesenteric artery occlusion, while the ischemic preconditioning group received 10 min of short ischemia and reperfusion before 45 min of prolonged occlusion. A cell counter was used to analyze blood obtained from rats before and after the surgical procedures and the hematological results were compared among the groups. RESULTS: The results showed significant differences in hematimetric parameters among the groups. The parameters that showed significant differences included lymphocyte, white blood cells and granulocyte counts; hematocrit; mean corpuscular hemoglobin concentration; red cell deviation width; platelet count; mean platelet volume; plateletcrit and platelet distribution width. CONCLUSION: The most remarkable parameters were those related to leukocytes and platelets. Some of the data, including the lymphocyte and granulocytes counts, suggest that ischemic preconditioning attenuates the effect of intestinal ischemia/reperfusion on circulating blood cells. Our work contributes to a better understanding of the hematological responses after intestinal ischemia/reperfusion and IPC, and the present findings may also be used as predictive values. .

Detección de la mutación V617F del genJAK2 mediante análisis de disociación de alta resolución / Detection of V617F mutation of JAK2 gene by high resolution melting / Detecção da mutação V617F do gene JAK2 através da análise de dissociação em alta resolução

La Policitemia Vera (PV), la Trombocitemia Esencial (TE) y la Mielofibrosis Primaria (MP) son neoplasias mieloproliferativas caracterizadas por una proliferación excesiva de una o más líneas mieloides. En el año 2005 se identificó una mutación somática en el gen Janus kinase 2 (JAK2), que resulta en el reemplazo en la proteína de una fenilalanina por una valina en la posición 617 (JAK2 V617F). Esta mutación se encuentra en el 95% de pacientes con PV, y en la mitad de los casos de TE o MP. Se han descripto metodologías que permiten identificar esta mutación: dentro de las más utilizadas se encuentran la ARMS PCR (del inglés Amplification Refractory Mutation System PCR), la secuenciación y recientemente la HRM (High Resolution Melting). En este trabajo se estudió la detección de JAK2 V617F en muestras de pacientes con desórdenes mieloproliferativos mediante HRM, determinando especificidad y sensibilidad de la misma, comparándola con la ARMS PCR y la secuenciación. Los resultados demostraron que la técnica de HRM es superior a la secuenciación y equivalente a la ARMS PCR. Las metodologías sensibles y específicas para la detección de JAK2 V617F, en pacientes con neoplasias mieloproliferativas, son de gran importancia a nivel diagnóstico ya que permiten diferenciar entre desórdenes neoplásicos y condiciones reactivas.

Polycythemia Vera (PV), Essential Thrombocythemia (TE) and Primary Myelofibrosis (MP) are myeloproliferative neoplasias characterized by excessive proliferation of one or more myeloid lines. In 2005, a somatic mutation was identified in the Janus kinase 2 gene (JAK2), resulting in the replacement of a phenylalanine in the protein for a valine at position 617 (JAK2 V617F). This mutation was found in 95% of patients with PV, and in half of the cases of TE or MP. Different methodologies have been described to identify this mutation: the most used are ARMS PCR (Amplification Refractory Mutation System PCR), sequencing and recently HRM (High Resolution Melting). In this work, detection of JAK2 V617F was studied in samples from patients with myeloproliferative disorders using HRM, determining its specificity and sensitivity in comparison with ARMS PCR and sequencing. The results showed that the technique is superior to sequencing and equivalent to ARMS PCR. Sensitive and specific methodologies for the detection of JAK2 V617F in patients with myeloproliferative neoplasias are of great importance at diagnostic level since they can differentiate between neoplastic disorders and reactive conditions.

Policitemia Vera (PV), Trombocitemia Essencial (TE) e Mielofibrose Primária (MP) são neoplasias mieloproliferativas caracterizadas por uma proliferação excessiva de uma ou mais linhas mieloides. Em 2005, uma mutação somática foi identificada no gene Janus quinase 2 (JAK2), resultando na substituição na proteína de uma fenilalanina para uma valina na posição 617 (JAK2 V617F). Esta mutação é encontrada em 95% dos pacientes com policitemia vera, e na metade dos casos de TE ou MP. Foram descritas metodologias para identificar esta mutação: dentre as mais utilizadas estão ARMS PCR (Amplification Refractory Mutation System PCR), sequenciamento e, recentemente, HRM (High Resolution Melting). Neste trabalho a detecção de JAK2 V617F foi estudada em amostras de pacientes com neoplasias mieloproliferativas usando HRM, determinando a sensibilidade e especificidade da mesma, comparando-a com a ARMS PCR e o sequenciamento. Os resultados mostraram que a técnica de HRM é superior ao sequenciamento e equivalente à ARMS PCR. Sensíveis e específicas para a detecção de JAK2 V617F em pacientes com neoplasias mieloproliferativas, as metodologias são de grande importância em nível de diagnóstico, uma vez que permitem diferenciar entre doenças neoplásicas e condições reativas.

Tuberculin skin test and interferon-gamma release assay values are associated with antimicrobial peptides expression in polymorphonuclear cells during latent tuberculous infection

It has been reported that patients with progressive tuberculosis (TB) express abundant amounts of the antimicrobial peptides (AMPs) cathelicidin (LL-37) and human neutrophil peptide-1 (HNP-1) in circulating cells, whereas latent TB infected donors showed no differences when compared with purified protein derivative (PPD) and QuantiFERON®-TB Gold (QFT)-healthy individuals. The aim of this study was to determine whether LL-37 and HNP-1 production correlates with higher tuberculin skin test (TST) and QFT values in TB household contacts. Twenty-six TB household contact individuals between 26-58 years old TST and QFT positive with at last two years of latent TB infection were recruited. AMPs production by polymorphonuclear cells was determined by flow cytometry and correlation between TST and QFT values was analysed. Our results showed that there is a positive correlation between levels of HNP-1 and LL-37 production with reactivity to TST and/or QFT levels. This preliminary study suggests the potential use of the expression levels of these peptides as biomarkers for progression in latent infected individuals.

Hematology of Swiss mice (Mus musculus) of both genders and different ages

PURPOSE: To describe the hematologic values of male and female, young and adult, Swiss mice (Mus musculus). METHODS: Mus musculus (n=14) were randomly selected and separated by gender. The male and female, young and adult animals were sedation to obtain a blood sample, by intracardiac route at 30, 45, 60, 75, 90, 105 and 120 days after birth. RESULTS: The Swiss mouse hemogram values obtained, in relation to total eosinophils, basophils, and number of platelets, there was no statistical differences according to the genders or the age of the animals. Regarding the erythrocyte, hemoglobin and hematocrit values obtained, these were higher in females. The RDW-CD and MPV values were higher in the females than in the males. CONCLUSIONS: Lymphocytes are the predominant cells in the peripheral blood. The collection of 800 µL of blood by intracardiac route, every 15 days, did not affect the health of the animals. Analyses of the blood samples contribute to the experimental models provided by the Central Animal Facility of UFMS and used by professors. .